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Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens.
J Med Microbiol. 2004 Jul; 53(Pt 7):603-607.JM

Abstract

Between January 2002 and July 2003, 173 bronchoalveolar lavage (BAL) specimens from 150 patients (19 HIV-infected and 131 non-HIV-infected patients) were evaluated for identification of Pneumocystis jiroveci (formerly known as Pneumocystis carinii f. sp. hominis) using staining techniques, conventional PCR (mtLSUrRNA gene) and real-time PCR (MSG gene). Test results were compared to Pneumocystis pneumonia (PCP) confirmed by typical clinical findings and response to treatment. Sensitivity and specificity of the techniques were 60 and 100% for staining (where either one or both techniques were positive), 100 and 87.0% for conventional PCR and 100 and 84.9 % for real-time PCR, respectively. The use of a concentration of 10(3) copies of DNA per capillary of BAL as a cut-off (determined by real-time PCR) increased specificity from 84.9 to 98.6% without reducing the sensitivity of the technique. This technique is rapid (<3 h) and therefore of major interest in differentiating between asymptomatic carriage and PCP. A BAL specimen with <10(3) copies per capillary of Pneumocystis-specific DNA is more likely to indicate a chronic carrier state, but in such cases follow-up is required to ensure that the patient is not in the early stage of an active PCP.

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Authors+Show Affiliations

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

Laboratory of Parasitology and Mycology, Hôpital Nord1 and Department of Infectious and Tropical Diseases, Hôpital Bellevue2, University Hospital of Saint Etienne, 42055 Saint Etienne, France.

MeSH

AIDS-Related Opportunistic InfectionsBronchoalveolar Lavage FluidCarrier StateDNA, FungalFalse Negative ReactionsFalse Positive ReactionsHumansImmunocompromised HostPneumocystis cariniiPneumonia, PneumocystisPolymerase Chain ReactionSensitivity and SpecificityStaining and Labeling

Pub Type(s)

Comparative Study

Evaluation Study

Journal Article

Language

eng

PubMed ID

15184529

Citation

Flori, Pierre, et al. "Comparison Between Real-time PCR, Conventional PCR and Different Staining Techniques for Diagnosing Pneumocystis Jiroveci Pneumonia From Bronchoalveolar Lavage Specimens." Journal of Medical Microbiology, vol. 53, no. Pt 7, 2004, pp. 603-607.

Flori P, Bellete B, Durand F, et al. Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens. J Med Microbiol. 2004;53(Pt 7):603-607.

Flori, P., Bellete, B., Durand, F., Raberin, H., Cazorla, C., Hafid, J., Lucht, F., & Sung, R. T. M. (2004). Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens. Journal of Medical Microbiology, 53(Pt 7), 603-607. https://doi.org/10.1099/jmm.0.45528-0

Flori P, et al. Comparison Between Real-time PCR, Conventional PCR and Different Staining Techniques for Diagnosing Pneumocystis Jiroveci Pneumonia From Bronchoalveolar Lavage Specimens. J Med Microbiol. 2004;53(Pt 7):603-607. PubMed PMID: 15184529.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens. AU - Flori,Pierre, AU - Bellete,Bahrie, AU - Durand,Fabrice, AU - Raberin,Hélène, AU - Cazorla,Céline, AU - Hafid,Jamal, AU - Lucht,Frédéric, AU - Sung,Roger Tran Manh, PY - 2004/6/9/pubmed PY - 2004/8/3/medline PY - 2004/6/9/entrez SP - 603 EP - 607 JF - Journal of medical microbiology JO - J Med Microbiol VL - 53 IS - Pt 7 N2 - Between January 2002 and July 2003, 173 bronchoalveolar lavage (BAL) specimens from 150 patients (19 HIV-infected and 131 non-HIV-infected patients) were evaluated for identification of Pneumocystis jiroveci (formerly known as Pneumocystis carinii f. sp. hominis) using staining techniques, conventional PCR (mtLSUrRNA gene) and real-time PCR (MSG gene). Test results were compared to Pneumocystis pneumonia (PCP) confirmed by typical clinical findings and response to treatment. Sensitivity and specificity of the techniques were 60 and 100% for staining (where either one or both techniques were positive), 100 and 87.0% for conventional PCR and 100 and 84.9 % for real-time PCR, respectively. The use of a concentration of 10(3) copies of DNA per capillary of BAL as a cut-off (determined by real-time PCR) increased specificity from 84.9 to 98.6% without reducing the sensitivity of the technique. This technique is rapid (<3 h) and therefore of major interest in differentiating between asymptomatic carriage and PCP. A BAL specimen with <10(3) copies per capillary of Pneumocystis-specific DNA is more likely to indicate a chronic carrier state, but in such cases follow-up is required to ensure that the patient is not in the early stage of an active PCP. SN - 0022-2615 UR - https://www.unboundmedicine.com/medline/citation/15184529/Comparison_between_real_time_PCR_conventional_PCR_and_different_staining_techniques_for_diagnosing_Pneumocystis_jiroveci_pneumonia_from_bronchoalveolar_lavage_specimens_ DB - PRIME DP - Unbound Medicine ER -